201247681 六、發明說明： 【發明所屬之技術領域】 本發明係有關N-节酿基-十字孢靈素（staur〇sp〇rine)的新 顆結晶形Π;製備N_节醯基_十字？包靈素的新賴、结晶形_ 方法包含N·节醯基·十字孢靈素的新賴結晶形π的組合 物，和Ν-苄醯基-十字孢靈素新穎結晶在溫血動物（特 別是人類）之診斷方法或治療方面的用途。 本發明係有關非晶形的苄醯基_十字孢靈素；製備非 晶形Ν-节醢基·十字孢靈素的方法，包含非晶形Ν_节醯基_ 十字孢靈素的組合物，和非晶形Ν_苄醯基_十字孢靈素在 /JEL血動物（特別疋人類）之診斷方法或治療方面的用途。 【先前技術】 藥品，Ν-苄醯基_十字孢靈素被用來作為一種抗腫瘤 劑。Ν-苄醯基-十字孢靈素的製備方法一般是業界已知 的。然而，亦已知的是，相同藥品之不同多晶型在某些具 醫學重要性的性質上確有大幅的差異。因此，Ν_苄醯基_ 十字孢靈素新穎的固態型和新的製備方法是持續需要的。 【發明内容】 根據某一態樣，本發明係提供…苄醯基_十字孢靈素的 結晶形II。Ν-苄醯基-十字孢靈素結晶形^^的尽射線繞射光 譜如圖1所描繪在反射角20為8.7±〇 2之處具有一譜峰。該 Ν-节醯基-十字孢靈素的結晶形11在95%的相對濕度和25。〇 下仍保持為乾燥的。 根據另一態樣，本發明係提供包含固態之Ν_苄醯基_十 165155.doc 201247681 字抱靈素的組合物，其中至少8G重量％的該固態ν·节醯基_ 十字抱靈素為X·射線繞射光譜如圖_描繪在反射角^為 8.7±0·2之處具有—错峰的結晶形π ^亦提供各式各樣的具 體實施例和變體。 根據另-態樣，本發明係提供包含Ν_¥醯基_十字抱靈 素的結晶形II和醫藥上可接受載體或稀釋劑的醫藥組合 物。該醫藥組合物較佳為經口服投藥者。 根據又另一態樣，本發明亦係有關非晶形的Ν_苄醯基_ 十字孢靈素，製備非晶形Ν_苄醯基_十字孢靈素的方法， 和包含非晶形Ν-苄醯基-十字孢靈素的組合物。 根據另一態樣，本發明係提供一種醫藥組合物，包括一 疾病預防上或醫療上有效量的Ν_苄醯基_十字孢靈素和一 或多種的醫藥上可接受賦形劑。本發明此態樣的醫藥組合 物可調配成例如經口服投藥者β。 根據又一態樣，本發明係提供純化十字孢靈素的方法。 根據另一態樣，本發明係提供令十字孢靈素和苄酐反應 製備Ν-节酿基-十字孢靈素的方法。此方法視情況進一步 包含播種该反應溶液，以製造非晶形的Ν_苄醯基—十字孢 靈素或Ν-苄醢基-十字孢靈素的結晶形π。 根據又一態樣，本發明係提供製備非晶形Ν_苄醯基-十 字孢靈素的方法，涉及喷霧乾燥包含Ν_苄醯基_十字孢靈 素的溶液。 亦提供包括以所述方法製得之疾病預防上或醫療上有效 量的Ν-苄醯基-十字孢靈素和一或多種的醫藥上可接受賦 165155.doc 201247681 形劑的醫藥組合物。 根據另—態樣’本發明係、提供製備N·㈣基·十字抱靈 素結晶形II的方法，該方法包含： ⑷提供料質子性或非質子性溶劑中的醯基_十字 孢靈素溶液； (b) 播種N-苄醯基-十字孢靈音 于把靈素的結晶形„，並讓該反應 混合物和一種醇溶劑接觸，形 啁形成一種沉澱物：和 (c) 分離此沉澱物，其乃n筚疏贫.. 八乃醯基-十字孢靈素的結晶形 II ° 也提供备式各樣的具體實施例和變體。 【實施方式】 除非另有定義’否則本文所用的所有技藝和科學術1 和本發明有關的一般技術者所公認的定義是 任何類似或等同於本文所述的方 k 貝白J用於本發日/ 的貫施和試射，㈣料㈣㈣的枝和 對本發明目的而言，以下術語係定義如下。 結晶化合物，本文指定為’’、结晶形n"且於下 N-节醯基_十字孢靈素的結晶形 ' 巾稱之為 射，和/或紅外線光譜學作鏜定。過X_射線的粉末續 種通常是無毒以及 包括對獸醫的使用 "醫藥上可接受”意指其有助於製備— 不是生物學上不受歡迎的醫藥組合物， 和/或八類醫學上的使用是可接受的。 "反溶劑"是當矣添加至某 該物質的溶解度的溶劑。 一物質的現有溶液中時會降低 165155.doc 201247681 ••組合物"乙詞包括（但不限於）粉末，溶液，懸浮液，凝 膠’軟膏，乳劑和/或其混合物。術語組合物意涵蓋包含 特定量之特定成份的產物，以及任何直接或間接導致特定 成份以特定量結合的產物。，，組合物"可包括一種單一化合 物或化合物的混合物。”化合物”是一種包括相同化學結構 之分子的化學物質。 術語”醫藥組合物"意涵蓋包含活性成份，構成載體之醫 藥上可接受賦形劑的產物，以及任何直接或間接導致兩種 或多種該等組份的結合，錯合或聚集的產物，或導致一或 多種該等組份分解的產物，或導致一或多種該等組份之其 他類型的反應或相互作用的產物。因此，本發明的醫藥組 合物涵蓋任何藉摻合活性成份，額外的活性成份和醫藥上 可接受賦形劑的組合物。 術語"賦形劑"意指醫藥組合物中不是活性成份（像是填 料，稀釋劑和載體）的組份^有助於製備醫藥組合物的賦 形劑通常是安全，無毒且既不是生物學上不受歡迎也不是 其他方面不受歡迎的，並且對獸醫的使用和人類醫學上的 使用疋可接受的。說明書和申請專利範圍中所用"醫藥上 可接受賦形劑"包括一和一種以上的此種賦形劑。 醫療上有效量"意指當投藥供治療或預防一種疾病之化 合物的量足以對這個疾病的治療或預防產生效果的量。 醫療上有效量會依化合物，待治療病患的疾病和其嚴重 性與年齡，體重等等而變。 當指的是一種化學反應時，術語"處理，，，，，接觸"和"反應,, 165155.doc 201247681 或1人疋可以交替使用的，並且指的是在適當條件下添加 ^口兩種或兩種以上的試劑’以製得所指和/或所希的 。應理解地是’產生所指和/或所希產物的反應未必 起因於-開始所添加之兩種試劑的結合，也就是說， ^可能是-或一種以上於混合物中生成的中間物，其最後 導致形成所指和/或所希的產物。 本文所用有關組合物的術語"大體上不含"意指該組合物 不3的物形無法以熟悉本技術者已知的方法偵測出。 術》。非晶形"意指大體上不含結晶形雜質的材料，或是 包含大量之結晶形雜質者。包含大量為雜質之結晶形質的 非曰曰形材料在此稱之為"基本上是非晶形的N-苄醯基·十字 忆靈素&圖4所示’結晶形雜質的存在不會產生一種對 大體上不含結晶材料的非晶形為典型之暈圈般的漂亮圖 案大體上不含結晶形的非晶形材料稱之為"N_苄醯基-十 子孢靈素的純非晶形"。圖3說明此形態的χ_射線繞射光譜 圖:此處圖案的暈圈形說明實質上不存在結晶構造。譜峰 在这些是結晶形特徵的區域内尤其是找不到的。 實例進—步說明獲得"基本上是非晶形的N_节醯基-十字 孢靈素"和"N_苄醯基-十字孢靈素的純非晶形"兩者的方 法。 N节酿基·十字孢靈素已知為（9α，10β，11β，13〇〇_Ν (2,3’1〇，11，12，13-六氫-1〇_曱氧基-9-甲基-1-氧-9，13_環氧_ lh，9H_ 一吲哚[1，2,3-gh:3，，2，，l'-lmp 比咯[3,4-j][l，7]苯并重 氮寧-11-基）甲基-苯甲醯胺）並具有以下的化學結構： I65155.doc 201247681201247681 VI. Description of the Invention: [Technical Field to Be Invented by the Invention] The present invention relates to a new crystalline form of N-synthesis-staur〇sp〇rine; preparation of N_ganglyl_cross? The new crystallization of the enantiomers, the crystalline form _ The method consists of a composition of N. sulphonyl·spironosin, a new crystalline form of π, and a novel crystal of Ν-benzyl hydrazino-craspellin in warm-blooded animals ( In particular, the diagnostic method or therapeutic use of humans. The present invention relates to an amorphous form of benzindenyl-spironosin; a process for preparing an amorphous form of Ν-醢 醢 · 十字 十字 十字 ,, comprising an amorphous Ν 醯 醯 _ 十字 十字 十字 十字 十字 的 的 的Use of amorphous Ν-benzyl hydrazino _ sporosporin in the diagnostic method or treatment of /JEL blood animals (especially scorpion humans). [Prior Art] The drug, Ν-benzyl hydrazine _ sporosporin was used as an antitumor agent. The preparation of bismuth-benzyl-based sporosporin is generally known in the art. However, it is also known that different polymorphs of the same drug do vary greatly in certain medically important properties. Therefore, the novel solid state and new preparation methods of Ν_benzyl hydrazino _ sporosporin are continuously required. SUMMARY OF THE INVENTION According to a certain aspect, the present invention provides a crystalline form II of benzinyl-streprise. The ray diffraction spectrum of the Ν-benzyl sulfhydryl-craspellin crystal form has a peak at a reflection angle of 20 8.7 ± 〇 2 as depicted in FIG. The crystalline form 11 of the Ν-knotyl-based sporosporin is at 95% relative humidity and 25. The underarm remains dry. According to another aspect, the present invention provides a composition comprising a solid Ν 醯 醯 醯 _ 165 165 155. doc 201247681 vocabulin, wherein at least 8 G weight % of the solid ν 醯 _ _ 十字 抱 抱The X-ray diffraction spectrum as shown in Fig. _ depicts a crystal form π^ having a --peak peak at a reflection angle of 8.7 ± 0. 2 also provides a variety of specific embodiments and variants. According to another aspect, the present invention provides a pharmaceutical composition comprising crystalline form II of Ν_¥醯基_十字抱灵素 and a pharmaceutically acceptable carrier or diluent. The pharmaceutical composition is preferably administered orally. According to still another aspect, the present invention is also directed to a method for preparing an amorphous Ν-benzyl hydrazino _ sporosporin, and an amorphous Ν-benzyl hydrazine. A composition of basal-aspirin. According to another aspect, the present invention provides a pharmaceutical composition comprising a disease prophylactically or medically effective amount of Ν-benzyl hydrazine-ross sporogenin and one or more pharmaceutically acceptable excipients. The pharmaceutical composition of this aspect of the invention can be formulated, for example, as an orally administered drug. According to still another aspect, the invention provides a method of purifying sporosporin. According to another aspect, the present invention provides a process for the preparation of sputum-frozen-spirulinin by reacting staurosporin with benzyl anhydride. The method further comprises, as the case may be, sowing the reaction solution to produce a crystalline form π of amorphous Ν-benzyl hydrazino-aspartate or Ν-benzyl hydrazino-craspellin. According to still another aspect, the present invention provides a process for the preparation of an amorphous bismuth-benzylidene-sporin, relating to spray drying a solution comprising Ν-benzyl hydrazino-tisporin. Also provided are pharmaceutical compositions comprising a prophylactically or medically effective amount of a disease-producing or medically effective amount of bismuth-benzyl-based sporosporin and one or more pharmaceutically acceptable agents 165155.doc 201247681. According to another aspect, the invention provides a method for preparing N. (di)-based spirulina crystal form II, the method comprising: (4) providing a sulfhydryl-sporosporin in a protic or aprotic solvent a solution; (b) seeding N-benzyl sulfhydryl-craspellin to crystallize the morpheme, and contacting the reaction mixture with an alcohol solvent to form a precipitate: and (c) separating the precipitate The substance, which is n筚-poor.. The crystalline form of octadecyl-craspellin II ° also provides a variety of specific examples and variants. [Embodiment] Unless otherwise defined, otherwise used herein All of the art and science 1 and the definitions recognized by the general practitioners of the present invention are any similar or equivalent to the square k-white J used for the present day/of the test and test, (4) material (four) (four) Branches and for the purposes of the present invention, the following terms are defined as follows: Crystalline compounds, designated herein as '', crystalline n" and in the crystalline form of the lower N-nodal thiol-spironosin' And/or infrared spectroscopy for determination. X-ray powder continued Species are generally non-toxic and include the use of veterinarians "pharmaceutically acceptable" means that they aid in the preparation - not biologically undesirable pharmaceutical compositions, and / or eight types of medical use are acceptable . "antisolvent" is a solvent that is added to the solubility of a substance. A substance may be reduced in an existing solution. 165155.doc 201247681 ••Composition" B includes, but is not limited to, powders, solutions, suspensions, gels, ointments, emulsions and/or mixtures thereof. The term composition is meant to encompass a product comprising a particular amount of a particular component, as well as any product that directly or indirectly results in the binding of a particular component in a particular amount. , the composition " may comprise a single compound or a mixture of compounds. A "compound" is a chemical that includes molecules of the same chemical structure. The term "pharmaceutical composition" is intended to cover the product comprising the active ingredient, the pharmaceutically acceptable excipients constituting the carrier, and any product which directly or indirectly causes the binding, mismatch or aggregation of two or more such components, Or a product which results in the decomposition of one or more of such components, or a product which results in one or more other types of reactions or interactions of such components. Thus, the pharmaceutical compositions of the present invention encompass any blended active ingredient, additional A combination of an active ingredient and a pharmaceutically acceptable excipient. The term "excipient" means a component of a pharmaceutical composition that is not an active ingredient (such as a filler, diluent, and carrier). Excipients for pharmaceutical compositions are generally safe, non-toxic and neither biologically undesirable nor otherwise undesirable, and are acceptable for veterinary use and human medical use. Instructions and patent applications "Pharmaceutically acceptable excipient" used in the scope includes one or more such excipients. Medically effective amount " means when administered for treatment The amount of a compound that prevents a disease from being effective in the treatment or prevention of the disease. The medically effective amount will vary depending on the compound, the condition of the patient to be treated and its severity and age, body weight, etc. When it is a chemical reaction, the terms "process,,,,,contact" and "reaction, 165155.doc 201247681 or one person can be used interchangeably, and refer to the addition of two types under appropriate conditions. Or two or more reagents' to produce the indicated and/or desired. It is understood that the reaction to produce the indicated and/or the desired product does not necessarily result from the combination of the two reagents added at the beginning. That is, ^ may be - or more than one intermediate formed in the mixture, which ultimately results in the formation of the indicated and/or desired product. The term "substantially free" in relation to the composition as used herein means The shape of the composition is not detectable by a method known to those skilled in the art. "Amorphous" means a material which is substantially free of crystalline impurities or contains a large amount of crystalline impurities. Including large The non-曰曰-shaped material which is a crystalline form of impurities is referred to herein as "substantially amorphous N-benzyl fluorenyl-cross-reagent& The presence of the 'crystalline impurity' shown in Fig. 4 does not produce a pair An amorphous form that is substantially free of crystalline material is a typical halo-like beautiful pattern. The amorphous material that is substantially free of crystalline form is called "N-benzylidene-triosporin pure amorphous" Fig. 3 illustrates a χ-ray diffraction spectrum of this form: the halo shape of the pattern herein indicates that there is substantially no crystal structure. The peaks are not particularly found in the regions where these are crystalline features. - Steps to obtain a method of obtaining both "substantially amorphous N_nodal thio-sporosporin" and "N_benzylidene-formin spores pure amorphous". N-flavored base · staurosporin is known as (9α, 10β, 11β, 13〇〇_Ν (2,3'1〇,11,12,13-hexahydro-1〇_曱oxy-9- Methyl-1-oxo-9,13_epoxy_lh,9H_ 一吲哚[1,2,3-gh:3,,2,,l'-lmp ratio [3,4-j][l , 7] benzodiazepine-11-yl)methyl-benzamide) and has the following chemical structure: I65155.doc 201247681
美國專利第5,093,330號（，33 0專利案）（該案之全文以弓丨用 的方式併入本文中）請求N_苄醯基-十字孢靈素。本發明特 定係有關如上所述下文稱之為式⑴化合物之沁苄醯基、十 字抱靈素衍生物之結晶形Π的特殊形態，以及有關N_节峨 基-十字孢靈素的非晶形。 不同固形的相同藥品也許具有不同的性質，包括其等作 為藥品之用途具機能含意的特徵在具醫學重要性的性質方 面（如溶解速率和生物可利用性）可能有报大的差異。同樣 地，不同的多晶形物可能會有不同的加工性質，像是吸溼 性’流動性料’其可能影響料商t製造用之活性醫藥 品的溶解度。 利用Bragg-Brenuno的對交幾何學，在以丨加吨mc χ i 上測量X_射線的粉末繞射光譜。圖•射線繞射光譜總 結於表1中。 I65l55.doc 201247681 表1·Ν-苄醯基-十字孢靈素之結晶形η改質的粉末狀χ射線 繞射譜峰 2Θ(度數) 3.4 6.0 7.8 8.7 9.2 9.7 10.1 10.4 11.2 12.6 14.1 15.7 16.8 18.2 18.9 19.3 19.6 20.2 24.5 d-間隔(埃) 相對強度 2 uk83u618590373976546 ,4π109·9·8·8·7·7·6·5·5·4·4·4·4·4'3 中中中強中中中低.中甲中中中中中中強強中 應謹記在心的是，觀察到的2Θ角或間隔值因所用的特定 繞射儀，化驗人員和樣品的製作技術的不同如有稍許的變 化是可預期的。相對譜峰強度有更多的變化也是可以預期 的。一化合物之精確結晶形的鑑定主要應以對相對譜峰強 度較不具重要性之觀察到的2Θ角為基礎。 本文記錄的每個2Θ角值會存在某誤差限度。一較佳變體 中，N-苄醯基-十字孢靈素結晶形η的指定誤差限度對每— 譜峰而言約為±0.2。 Ν-苄醯基-十字孢靈素的結晶形^也可用紅外線光譜學描 續其特徵。結晶形II會表現出如圖2之紅外線（ir)光譜分析 的獨有吸收圖案。IR的光譜分析是於Bruker IFS_55上測量 165155.doc •9- 201247681 的。N-苄醯基-十字孢靈素的結晶形π具有特有和其他多晶 型相區別的吸收值，IR光譜分析上的譜峰位置為約789， 773 ， 743 ， 704 ， 1066 ， 1026 , 1458 , 1398 ， 1383 ， 1602，1577 ’ 1497，1627，1680，2934 和 3055 公分-1。本 文記錄的每個特定吸收值會存在某誤差限度。特定吸收值 的指定誤差限度在1900-800公分範圍内約為2公分·ι。 一或一種以上的物理性質和/或光譜性質可以是描繪Ν· 节醯基-十字孢靈素的結晶形或多晶型特徵的基礎。 本發明亦提供一種包含固態Ν_苄醯基_十字孢靈素的組 合物，其中組合物中固態Ν_苄醯基-十字孢靈素的至少8〇% 是結晶形II ^該組合物的較佳型態為在調配醫藥產品時適 合用來作為活性成份的Ν-苄醯基-十字孢靈素固體粉末。 組合物中固態Ν•苄醯基_十字孢靈素剩餘的部份，也就是 Ν_节酿基-十字孢靈素總重的20%或少於20%的部份可以是 例如其他結晶形的Ν_苄酿基-十字抱靈素。一特定具體實 施例中，該組合物包含佔組合物中固態Ν_苄醯基_十字孢 靈素總重至少90% Ν_苄醯基_十字孢靈素的結晶形π。另一 特定具體實施例中’該組合物包含佔組合物中固態Ν-苄醯 基-十字孢靈素總重至少95% Ν_苄醯基_十字孢靈素的結晶 形II。 另層面中’本發明亦提供純化十字孢靈素的方法。同 時本發明並不限定於任何特殊的理論，發明人發現一種 涉及以下步驟的方法： (a)在醇溶劑中提供十字孢靈素的懸浮液； 165l55.doc 201247681 (b)讓此料液和甲烧賴接觸獲得-種溶液； (C)再讓此溶液和三乙胺接觸；以及 (d)分離出產物。 該方法進一步包括在質子性或非質子性溶劑中清洗該產 。質子性溶劑的非限制性實例有乙醇，而非質子性溶劑 的非限制性實例為四氫呋喃(THF)。本發明亦提供一種方 法，用來： ⑷在質子性或非質子性溶劑中提供粗十字抱靈素的溶 (b) 用純的十字孢靈素播種該溶液； (c) 讓此反應混合物和一醇系.、交逾丨丨姐飽 畔系冷劑接觸形成一種沉澱物； 以及 (d)分離出產物，其係經純化的十字孢靈素。 原料經純化後，該物質可被轉換成N_节醯基·十字孢靈 素的結晶形II或非晶形的醯基_十字抱靈素。該物質可 被轉換成Ν-f醢基·十字抱靈素的結晶形1](，此方法包括： ⑷在-㈣溶财，像是乙料，讓切孢靈素和节酸 酐反應；以及 (b) 用Ν-节酿基十字抱靈素的結晶形Π播種該反應溶 液，以製得產物； (c) 分離從N-苄醯基-十字孢靈素結晶出的產物。 材料或可利用以下方式轉換成非晶形的N•节醯基_十字 孢靈素： ⑷在-溶射，像是乙醇或卿中，讓十字孢靈素和节 165155.doc 201247681 酸酐反應；以及 (b)用非晶形的N-苄醯基-十字孢靈素播種該反應，以製 得產物； 0)分離為非晶形N-苄醯基-十字孢靈素的產物。 產物或可進一步用溶劑清洗多次以製得基本上為非晶 形的N-苄醯基-十字孢靈素。 喷霧乾燥的技術也可用來製備純粹為非晶形的N_节醯 基-十字孢靈素。U.S. Patent No. 5,093,330 (the <RTIgt The present invention specifically relates to a specific form of a crystalline form of a quinone-based group, a spirulina-like derivative, which is hereinafter referred to as a compound of the formula (1), and an amorphous form of N_cathene-based sporosporin. . The same drug with different solid forms may have different properties, including its functionally useful characteristics as a drug for medically important properties (such as dissolution rate and bioavailability) that may have large differences. Similarly, different polymorphs may have different processing properties, such as hygroscopic 'flowing materials' which may affect the solubility of the active pharmaceutical product used in the manufacture of the material. Using the orthogonal geometry of Bragg-Brenuno, the powder diffraction spectrum of X-rays was measured on a ton plus mc χ i . The graph ray diffraction spectra are summarized in Table 1. I65l55.doc 201247681 Table 1. Ν-Benzyl fluorenyl-craspellin crystal form η modified powdered χ ray diffraction peak 2 Θ (degrees) 3.4 6.0 7.8 8.7 9.2 9.7 10.1 10.4 11.2 12.6 14.1 15.7 16.8 18.2 18.9 19.3 19.6 20.2 24.5 d-interval (Angstrom) Relative intensity 2 uk83u618590373976546, 4π109·9·8·8·7·7·6·5·5·4·4·4·4·4'3 中中中强中Medium, medium and low. Medium, middle, middle, middle and strong strengths should be kept in mind. The observed 2 Θ angle or interval values are slightly different depending on the specific diffractometer used, the laboratory technician and the sample making technique. The changes are predictable. More variations in relative peak intensity are also expected. The identification of the precise crystal form of a compound should be based primarily on the observed 2 Θ angles that are less important for relative peak intensity. There is some margin of error for each 2 Θ angle value recorded in this paper. In a preferred variant, the specified error margin for the N-benzyl hydrazino-Casprin crystal form η is about ± 0.2 for each peak. The crystalline form of Ν-benzyl hydrazino-craspellin can also be characterized by infrared spectroscopy. Crystalline II exhibits a unique absorption pattern as shown by the infrared (ir) spectrum of Figure 2. IR spectral analysis was performed on a Bruker IFS_55 165155.doc •9-201247681. The crystalline form π of N-benzyl hydrazino-craspellin has an absorption value unique to other polymorphs, and the peak positions on the IR spectrum are about 789, 773, 743, 704, 1066, 1026, 1458. , 1398, 1383, 1602, 1577 '1497, 1627, 1680, 2934 and 3055 cm-1. There is a margin of error for each specific absorption value recorded in this document. The specified error limit for a particular absorption value is approximately 2 cm·ι in the range of 1900-800 cm. One or more physical properties and/or spectral properties may be the basis for characterizing the crystalline or polymorphic character of the Ν·醯 醯-PS-sporospin. The present invention also provides a composition comprising a solid Ν-benzyl hydrazino-disclostatin, wherein at least 8 % of the solid Ν-benzyl hydrazino-ross sporogenin in the composition is a crystalline form II ^ A preferred form is a bismuth-benzyl-based spironolactin solid powder suitable for use as an active ingredient in the formulation of a pharmaceutical product. The remaining portion of the solid quinone-benzylidene _ sporosporin in the composition, that is, 20% or less than 20% of the total weight of the 节 节 节 - 十字 十字 十字 十字 可以 可以 可以 可以 可以 可以Ν 苄 benzyl ketone - cross agave. In a specific embodiment, the composition comprises at least 90% of the total weight of the solid Ν-benzyl hydrazino _ sporosporin in the composition. In another specific embodiment, the composition comprises at least 95% of the total weight of the solid quinone-benzyl hydrazino-ross sporogenin in the composition. In another aspect, the invention also provides a method of purifying sporosporin. While the invention is not limited to any particular theory, the inventors have discovered a method involving the following steps: (a) providing a suspension of staurosporine in an alcohol solvent; 165l55.doc 201247681 (b) letting this liquid and The solution is contacted with a solution; (C) the solution is contacted with triethylamine; and (d) the product is isolated. The method further comprises washing the product in a protic or aprotic solvent. A non-limiting example of a protic solvent is ethanol, and a non-limiting example of a non-protic solvent is tetrahydrofuran (THF). The invention also provides a method for: (4) providing a solution of crude cross-aglin in a protic or aprotic solvent; (b) seeding the solution with pure staurosporine; (c) allowing the reaction mixture and An alcohol system, a sputum-supplemented refrigerant is contacted to form a precipitate; and (d) a product is isolated, which is a purified staurosporine. After purification of the starting material, the material can be converted into crystalline form II of N_gangyl-based sporosporin or amorphous sulfhydryl-cross-agrow. The substance can be converted into a crystalline form of Ν-f 醢 · 十字 十字 灵 ( ( ( , , , ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( 以及 以及 以及 以及 以及 以及 以及 以及 以及 以及 以及 以及 以及(b) seeding the reaction solution with a crystalline form of Ν-节 基 十字 十字 十字 , , , , , , ; ; ; ; ; ; ; ; ; ; ; ; 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该 该Conversion to amorphous N• 醯 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ The amorphous N-benzyl hydrazino-craspellin is sown to produce the product; 0) is isolated as the product of amorphous N-benzyl hydrazino-craspellin. The product may be further washed a plurality of times with a solvent to produce a substantially amorphous N-benzyl hydrazino-craspellin. Spray drying techniques can also be used to prepare purely amorphous N-nodal thiol-ross sporogenin.
165155.doc 201247681 本發明也提供包含基本上為非晶形之队节酿基_十字抱 靈素的組合物。本發明亦提供包含純粹為非晶形之n•节酿 基-十字抱靈素的組合物。 亦提供製備N-节酿基-十字抱靈素之結晶形π的方法。該 方法涉及： ⑷在質子性或非質子性溶劑中提供酿基·十字抱靈 素的溶液； (b)用N-f醯基_十字抱靈素的結晶仙播種該溶液； ⑷讓此懸浮液和一醇系溶劑接觸形成一種沉澱物；以及 (句分離出產物，其祕节醯基-十字孢靈素的結晶形 II 〇 質子性或非質子性溶劑的非限制性實例列於下表中： 實例 节醇 乙醇 二甲亞砜（DMSO) 二曱醯胺（DMF)165155.doc 201247681 The present invention also provides compositions comprising substantially amorphous squash-based spirulina. The present invention also provides a composition comprising a purely amorphous n•throttle-cross-aggregin. A method of preparing the crystalline form π of N-flavoring-cross-agglomerating is also provided. The method involves: (4) providing a solution of a stilbene/cross agavein in a protic or aprotic solvent; (b) seeding the solution with a crystal scent of Nf thiol_cross-ocalin; (4) allowing the suspension and Contacting the monoalcoholic solvent to form a precipitate; and (for the separation of the product, the non-limiting examples of the crystalline form II protonic or aprotic solvent of the secretory thiol-spironosin are listed in the following table: Example phenol alcohol dimethyl sulfoxide (DMSO) diamine (DMF)
THF 乙酸 聚乙二醇（PEG 200) 從上面所指明溶劑所得結晶B„iX_射線粉末繞射圖的 譜峰強度為十到低，因為在晶格結構内存在不同的溶劑分 165155.doc 201247681 子其強度會有一些變化。 一具體實施例中’係跡节醯基-十字孢靈素溶解於节 醇中，透過添加N-节醯基·十字孢靈素結 晶形II的種晶而沉 澱，並添加乙醇作為反溶劑，以及獲得N-节醯基_十字孢 靈素的結晶形II。該方法具有高的再現性，並且所得結晶 產物有良好的過濾性質。B6中殘餘的溶劑量約為4 3重量/ 重量❶/。的节醇和0.5_1重量/重量％的乙醇。流程圖解如下：THF acetic acid polyethylene glycol (PEG 200) The peak intensity of the crystalline B„iX_ray powder diffraction pattern obtained from the solvent specified above is ten to low because there are different solvents in the lattice structure. 165155.doc 201247681 There are some changes in the intensity of the sub-segment. In a specific example, the 'stranded sulfhydryl-sporosporin is dissolved in the phenol, and precipitated by adding the seed crystal of the N-nodal thiol-aspirin crystal form II. And adding ethanol as an anti-solvent, and obtaining the crystal form II of N-nodal thiol_craspellin. The method has high reproducibility, and the obtained crystalline product has good filtration properties. The amount of residual solvent in B6 is about It is 43 wt/wt sterol and 0.5 _1 wt/wt ethanol. The flow chart is as follows:
IP-方法 B4 爷醇和乙醇中 的結晶反應IP-method B4 crystallization reaction in alcohol and ethanol
上面有關各別結晶形的選擇性製備條件並非決定性的。 例如其通常可以改變像是式（I)化合物相對溶劑和反溶劑重 量比的參數。 也提供包含N-苄醢基-十字孢靈素的結晶形„或非晶形N· 节醢基-十字孢靈素和醫藥上可接受載體的醫藥組合物。 除了活性化合物以外，該醫藥組合物另包括一或一種以上 的醫藥上可接受載體’亦已知為一般缺乏藥學活性但具有 165155.doc • 14 - 201247681 各種有用性質（例如可增加溶解度，殺菌力，生物可利用 性和容易使醫藥組合物調配之特性）的賦形齊卜這些載體 為醫藥上可接受者，代表其等當攝取適量時對人類或動物 是無害的並且和-既定配方中的其他成份是相容的。載體 可以疋固態，半固態或是液態，且可和大塊的化合物作調 配，但最終仍為單元劑量形的配方，也就是物理上是分離 的直到包含-特定量的活性成份，像是_或膠囊。該醫 藥組合物可包括除了本發明化合物以外，一或多種的活性 醫藥化合物。 該醫藥組合物可以是懸浮液，溶液，藥液酉也，氣溶谬 或固態的劑型。 該醫藥組合物可以是各種適合不同模式投藥的配方，包 括（但不限於）吸入，口服，直腸投藥，非經腸投藥（包括皮 下，皮内，肌内和靜脈内投藥），可植入投藥和經皮投 藥。-已知情況中，最適當的投藥途徑視所投藥對象的症 狀’所希的治療期間，待治療疾病的種類和嚴重性，以及 待使用的特殊酉己方而定。配方可以是散裝或為單位劑型 者，並且對一已知配方而言可以業界熟知的方法製備。 單位劑型中所包括之活性成份的量視其中所含活性成份 之配方的類型而定。醫藥組合物通常包括約〇1重量％至約 99重量。/。的活性成份’對口服投藥而言較佳為約i重”。至 約50重量。/。，而對非經腸投藥而言約〇2重量％至約π重曰 %。 ”、 置 適合口服投藥用的配方包括膠囊（硬式或軟式），扁囊 165155.doc 201247681 :活二糖毁，栓劑和藥片，每-種皆包含-已確定量 m:物；可為粉末或粒狀’水性或非水性液體中的 y杯：：：液’或是水包油或油包水的乳液。此等配方可 二二藥業的合適方法製備’包括將其和活性化合物與 、的載體或多種載體結合。每單位劑量固體配方中 所飞的量可如先前技藝中製備n•节醯基十字抱靈素 所述。 /發明的另-層面係提供使用該等化合物和本發明之醫 樂組合物的治療方法。本發明的化合物和組合物可將一用 來治療和預防，330專利中所揭示疾病所用之有效量投予一 對象。所謂的對象意指人類或動物，較佳為人類。本發明 議計的動物包括任何可以本發明化合物作安全治療的動 物。最顯著的是結晶形„和非晶形的㈣酿基十字抱靈素 因蛋白活化酵素C (PKC)的抑制作用而具有高抗增殖和抗 腫瘤的活性，其極適合用來治療癌症。再者，PKC的高選 ，性和有效的抑制作用對病患具有優良的臨床效果，也就 是延緩或壓制疾病的發展，是同樣可接受的攝生法。潛在 的應用包括各種的固體腫瘤’較特定而言例如是乳癌，結 腸癌，卵巢癌和血f此外，其他各種被PKC活性影㈣ 徵兆可以這些組合物作有效的治療，包括抗多藥性 (臟)，目前使用的一般癌症化療法和發炎性疾病最主要 的問題之一。 本發明特別係有關本文揭示用來治療該等疾狀一或用 來製備其治療用的藥學試劑的結晶形U和非晶形的N-节醯 165155.doc •16· 201247681 基-十字抱靈素。 本發明亦係有關治療罹患該等疾病之溫血動物的方法， 特別是腫瘤的疾病，其中係將某量（尤其是具有抗増殖和 抗腫瘤功效的量）的結晶形π或非晶形的N_苄醯基·十字抱 靈素（其可有效對抗有關的疾病）投予需要此一治療的溫血 動物。再者’本發明係有關結晶形13[或非晶形的N_苄醯基_ 十字孢靈素用於抑制上述PKC的用途，或用來製備可用以 治療人類或動物體的醫藥組合物，特別是用來治療各種固 體腫瘤，較特定而言例如是乳癌，結腸癌，印巢癌和血 癌。視人種，年齡，各別症狀，投藥模式和討論中的臨床 徵候而定，將有效劑量（例如曰劑量為約卜乃⑼毫克，較佳 為1-1000毫克，尤佳為5_500毫克）投予體重約7〇公斤的溫 血動物。 本發明亦係有關包含-有效量(尤其是預防或治療該等 疾病之一的有效量）之結晶形„或非晶形的式⑴N_节醯基_ 十字孢靈素’ it同醫藥上可接受載體的藥劑，此醫藥上可 接受載體適合於局部；冑内，例如口服或直腸或非經腸的 投藥並且可以是無機的或有機的和固體或液體的。包含活 性成份連㈣㈣（例如乳糖，㈣糖，隸，甘露糖 醇，山梨糖醇，纖維素和/或甘油；和/或潤滑劑，例如矽 石/月石’硬月曰酸或其鹽’典型為硬脂酸鎂或硬脂酸鈣； 和/或PEG)的鍵劑或明膠膠囊特別係用來供口服投藥用。 鍵劑可同樣_合劑’例如石夕銘酸鎮，澱粉，通常是玉 米’小麥或求澱粉，明膠，甲基纖維素，缓基甲基纖維素 165155.doc 201247681 鈉和/或聚乙烯基吡咯啶酮；以及如果真的想要的話，亦 包括崩解劑’例如澱粉，瓊脂，„酸或其鹽，通常為海 藻酸鈉，·和/或起泡的混合物，或是吸附劑，著色劑，調 味料和甜味劑。本發明具藥學活性的化合物可進一步以供 非經腸投藥的製劑或浸泡溶液的型態使用。此等溶液較佳 為等滲透壓的水溶液或懸浮&，這#可在使用前先行製 備，例如對包含活性成份（不論是單獨或和載體（例如甘露 糖醇）一起）的凍乾製劑而言。製藥物質可經消毒和/或可包 含賦形劑，例如防腐劑，安定劑，溼潤劑和/或乳化劑； 溶解劑；調節滲透壓的鹽；和/或緩衝液。本發明的藥學 製劑（如果真的想要的話，其可包含其他具藥學活性的2 質，像疋抗生素）以本身已知的方式製備，例如透過傳統 的混合，粒化，塗覆，溶解或凍乾方法，並包含約卜 1〇〇%(尤其是約1%至約20%)的多種或單一活性物質。 本發明進一步藉參考下列詳細說明本發明之化合物和組 :物的製備以及其等之用途的實例而定義。對熟悉此項技 術者而言，材料和方法的許多改變可在不偏離本發明之目 的和利益情況下得以實施是顯而易知的。後續的實例無音 用來限制本發明之範圍為上文所定義的範圍或如以下之申 請專利範圍所定義的範圍。 實例 實例1利用甲烷磺酸和三乙胺之十字孢靈素幻變為十字 孢靈素B2的純化方法 進料十字抱靈素B1和乙醇至一反應器中。加熱懸浮液至 165155.doc 201247681 約70 C。將曱院續酸加入該懸浮液中，並老化所得溶液。 添加活化的木炭和Hyflo ’並老化該混合物。過滤此懸浮 液’並用乙醇清洗該濾器裝置和濾餅多次。將清澈經過滤 的溶液移轉到另一反應器中，並冷卻至約6〇。〇。緩緩將於 乙醇中稀釋的三乙胺加入該反應混合物中。之後，冷卻此 懸浮液至約20°C並予以老化。十字孢靈素B2以過濾的方式 分離。用乙醇漂洗產物數次》 實例2透過結晶純化粗的十字孢靈素B1到十字孢靈素B2 進料十字孢靈素B 1和苄醇至一反應器中。加熱懸浮液至 約85°C。將活化的木炭和Hyflo加入該溶液中，並再次老 化該混合物。約9(TC的溫度下過濾此懸浮液。用节醇清洗 該濾器裝置和濾餅多次》將清澈經過濾的溶液移轉到另一 反應器中，並冷卻至約70°C。溶液以懸浮於乙醇中的十字 孢靈素B2播種。之後，將乙醇加入此懸浮液中並老化此懸 浮液’然後冷卻至約〇°C。約〇°C下再度老化該懸浮液至少 2小時。十字孢靈素B2以過濾的方式分離。用乙醇漂洗產 物數次。 實例3苄基化十字孢靈素B2以製備基本上是非晶形的节 醯基-十字孢靈素 進料含苄酸肝的十字抱靈素B1，乙醇和水至一反應器 中。加熱該反應混合物至約70°C。過濾、此溶液並用乙醇清 洗該濾器裝置和濾餅多次。將經過濾的溶液移轉到另—反 應器中，並冷卻至約601。緩緩地將水加入此溶液中。用 非晶形的N- >酿基-十字抱靈素播種。老化此懸浮液，然 165155.doc 201247681 後冷卻至約20°C，然後再予以老化。Ν·苄醯基-十字孢靈 素以過濾的方式分離。用純乙醇漂洗Ν-苄醯基-十字孢靈 素溼的濾餅數次。然後將溼的濾餅再懸浮於乙醇中的渡器 裝置中，約25°C下老化，然後冷卻至約〇-5°C，然後再予以 老化。重覆該方法步驟數次。N-苄酿基-十字孢靈素以過 濾的方式分離。在外部溫度為約40°C以及真空度為約ι〇_ 20毫巴的真空烘箱中乾燥此溼的濾餅約24小時，獲得產 物：35-40克的乾濾餅：27克的理論減輕重量。產率：相 對B1 100%為約82%的理論減輕值。 上述反應中的乙醇可以THF取代。 實例4苄基化十字孢靈素B2以製備結晶形的〜节醯基-十 字孢靈素B4 進料含苄酸酐的十字孢靈素B2，純乙醇和水至一反應器 中。加熱該反應混合物至約7 0。(3並予以老化。反應結束之 際，用懸浮於純乙醇中的N-苄醯基-十字孢靈素結晶形1；[播 種該反應混合物。老化後，緩緩地將純的乙醇和水加入此 懸浮液中。再度老化此懸浮液，然後在N_苄醯基_十字孢 靈素以過濾方式分離之前冷卻至約〇t：。用純的乙醇漂洗 N苄酿基-十子抱靈素溼的濾餅數次。然後置於外部溫度 為60°C以及真空度為約5_30毫巴的真空烘箱中乾燥約16小 時’獲得產物· 39.7G克，乾式〉慮餅：27·2克的理論減輕重 I。產率.相對Β1 1〇〇〇/0約9ΐ.5〇/〇的理論減輕值。 實例S利用苄基醇和乙醇製備結晶形11的Ν苄醯基十字 孢靈素 165155.doc •20- 201247681 進料粗的十予孢靈素和苄基醇至一反應器中。加熱該混 β物至85 C，並接著老化此溶液。冷卻溶液至約7〇。〇，然 後用懸浮於純乙醇中. 畔甲之N-V醯基_十字孢靈素則純的結晶 形II播種。之後’添加乙醇，並老化此懸浮液，然後在N_ 节酿基-十字孢靈素結晶形„以過濾、方式分離之前冷卻至約 〇°C。溼的濾餅用乙醇漂洗數次並乾燥之。 實例6利用乙睃和乙醇製備結晶形11的]^苄醯基十字孢 靈素 進料粗的十字孢靈素和乙酸至一反應器中。加熱該混合 物至約70°C，並接著老化此溶液。用懸浮於純乙醇中之N_ 苄醯基-十字孢靈素B6純的結晶形Π播種此溶液。之後，添 加乙醇至該懸浮液中，並老化此懸浮液，然後在Ν_苄醯 基-十字孢靈素結晶形II以過濾方式分離之前冷卻至約 20°C。溼的濾餅用乙醇漂洗數次並乾燥之。 實例7利用PEG製備N-苄醯基-十字孢靈素的結晶形η的 晶想 進料粗的Ν-苄酿基-十字孢靈素和peg至一反應器中。 加熱該混合物至約90°C，並接著老化此溶液。冷卻此溶液 至約70°C，然後用懸浮於純乙醇中之N-苄醯基-十字孢靈 素B6純的結晶形II播種。之後，緩緩添加比例為1 : 1的乙 醇和水，並在冷卻至20°C以前老化此懸浮液。以過濾方式 分離N-苄醯基-十字孢靈素的結晶形II。溼的濾餅用乙醇漂 洗數次並乾燥之。 實例8利用DMSO製備N-苄醯基-十字孢靈素的結晶形π 165155.doc -21 - 201247681 進料N-苄醯基-十字孢靈素和DMSO至一反應器中。加熱 該混合物至約7〇°C ’並接著老化此溶液，然後用懸浮於乙 醇中之N-节酿基-十字孢靈素B6純的結晶形II播種。之 後，添加乙醇至懸浮液中，並在70°C下老化此懸浮液，然 後冷卻至約2〇°C。以過濾方式分離N-苄醯基-十字孢靈素 的結晶形II以前再度老化此懸浮液。產物用乙醇漂洗數次 並乾燥之。 實例9利用DMF製備N-苄醯基-十字孢靈素的結晶形π 進料N-苄醯基-十字孢靈素和DMF至一反應器中。加熱 該混合物至約70°C，老化此溶液，然後用乙醇中之N-苄醯 基-十字孢靈素B6純的結晶形II播種。之後，添加乙醇至懸 浮液中，並在約70°C下老化此懸浮液，然後冷卻至約 20°C。以過濾方式分離N-苄醯基-十字孢靈素的結晶形Π以 前再度老化此懸浮液。產物用乙醇漂洗數次並乾燥之。 實例10利用DMF製備N-苄醯基-十字孢靈素的結晶形π 進料非晶形的N-苄醯基-十字孢靈素和THF至一反應器 中。加熱該混合物至約65°C，老化此溶液，然後用THF中 之N-苄醯基·十字孢靈素B6純的結晶形Π播種。老化此懸 浮液’然後冷卻至約2(TC，並以過濾方式分離N-苄醯基-十字孢靈素的結晶形II以前再度老化此懸浮液。產物用 THF漂洗數次並乾燥之。 實例11利用乙醇製備N-苄醯基-十字孢靈素的結晶形u 進料非晶形的N-节酿基-十字抱靈素和乙醇至一反應器 中。加熱該混合物至約7(TC，老化，然後用乙醇中之…苄 165155.doc •22· 201247681 醯基-十字孢靈素B6的結晶形II播種。老化此懸浮液，冷 卻至約20°C，並以過濾方式分離N-苄醯基-十字孢靈素的 結晶形Π以前再度老化此懸浮液。產物用乙醇漂洗數次並 乾燥之。 實例12喷霧乾燥實驗以獲得非晶形的PKC412 將10克的PKC412(結晶形II)溶解於160克的二氯曱烷中 以製備進料溶液。進料備妥後被原子化成液滴，有機溶劑 透過加熱的氮於乾室中蒸發出來，加熱的氮會導致生成非 晶形的PKC412。入口溫度設定為60°C，而出口溫度則設 定為40°C。喷霧乾燥後殘餘的溶劑量約為1%重量/重量。 實例13喷霧乾燥實驗以獲得非晶形的PKC412 將7克的PKC412(結晶形II)溶解於約400克的THF中以製 備進料溶液。進料備妥後被原子化成液滴，有機溶劑透過 加熱的氮於乾室中蒸發出來，加熱的氮會導致生成非晶形 的PKC412。入口溫度設定為110-120°C，而出口溫度則設 定為70-80°C。喷霧乾燥後殘餘的溶劑量約為3°/。重量/重 量。真空下進一步乾燥PKC412，獲得不含溶劑的材料。 實例14喷霧乾燥實驗以獲得非晶形的PKC412 將7克的PKC412(結晶形II)溶解於約350克70: 30重暈/重 量的乙醇/乙酸中以製備進料溶液。進料備妥後被原子化 成液滴，有機溶劑透過加熱的氮於乾室中蒸發出來，加熱 的氮會導致生成非晶形的PKC412。入口溫度設定為 215°C ’而出口溫度則設定為120°C。喷霧乾燥後殘餘的溶 劑量低於0.5%。不需要作進一步的傳統乾燥處理。 165155.doc -23- 201247681 實例15含N-苄醯基-十字孢靈素的結晶形II的錠劑 含100毫克標題中所命名之活性物質的錠劑通常是依下 列組成製備： 組合物含量 （毫克) 活性成份 100 結晶乳糖 240 Avicel 80 PVPPXL 20 一種高度分散的矽膠 2 硬脂酸鎂 5 總計 447 製備方法：活性物質和載體材料混合，並置於一壓錠機 (Korsch EKO，衝壓直徑為10毫米）上加以壓縮 Avicel是微晶纖維素（FMC，費城，美國） PVPPXL是聚乙烯基聚吡咯啶酮，經交聯的（巴地斯，德 國）The above selective preparation conditions for the respective crystal forms are not critical. For example, it is generally possible to vary the parameters of the compound of formula (I) relative to the solvent to antisolvent weight ratio. A pharmaceutical composition comprising a crystalline form of N-benzyl hydrazino-craspellin or an amorphous N. sulphonyl-sporosporin and a pharmaceutically acceptable carrier is also provided. In addition to the active compound, the pharmaceutical composition Also included is one or more pharmaceutically acceptable carriers that are also known to be generally devoid of pharmaceutically active but have various useful properties (eg, increased solubility, bactericidal power, bioavailability, and ease of medicinal properties) (165155.doc • 14 - 201247681) Characterization of the properties of the composition) These carriers are pharmaceutically acceptable, representing that they are not harmful to humans or animals when ingested in an appropriate amount and are compatible with other ingredients in a given formulation.疋 Solid, semi-solid or liquid, and can be formulated with bulk compounds, but ultimately still in the form of unit doses, that is, physically separated until containing a specific amount of active ingredients, such as _ or capsules The pharmaceutical composition may include one or more active pharmaceutical compounds in addition to the compound of the present invention. The pharmaceutical composition may be a suspension, a solution, or a drug. A pharmaceutical composition that can be administered in a variety of modes including, but not limited to, inhalation, oral, rectal administration, parenteral administration (including subcutaneous, intradermal, and intramuscular). Intravenous and intravenous administration), implantable administration and transdermal administration. - In the known case, the most appropriate route of administration depends on the symptom of the subject being administered, the type and severity of the disease to be treated, and The formulation may be in the form of a bulk or a unit dosage form, and may be prepared by a method well known in the art for a known formulation. The amount of the active ingredient included in the unit dosage form depends on the active ingredient contained therein. The type of the formulation depends on the pharmaceutical composition generally comprising from about 1% by weight to about 99% by weight of the active ingredient 'preferably about i by weight for oral administration." Up to about 50 weight. /. For parenteral administration, about 2% by weight to about π% by weight. Formulations suitable for oral administration include capsules (hard or soft), sac 165155.doc 201247681: live disaccharide, suppositories and tablets, each containing - a defined amount of m: substance; may be powder or A granule of 'y-cup:::liquid' in an aqueous or non-aqueous liquid or an oil-in-water or water-in-oil emulsion. These formulations can be prepared by a suitable method of the second pharmaceutical industry' including the active compound and The carrier or a plurality of carriers are combined. The amount of each unit dose of the solid formulation can be as described in the prior art for the preparation of n• 醯 十字 十字 十字 。 。. / The other layer of the invention provides the use of the compounds and the present invention Therapeutic methods of the medical therapeutic composition. The compounds and compositions of the present invention can be administered to a subject in an amount effective for the treatment and prevention of the diseases disclosed in the '330 patent. The so-called subject means human or animal, Preferably, the animal of the present invention includes any animal which can be safely treated by the compound of the present invention. The most remarkable is the inhibition of the crystalline form and the amorphous (four) stimulating-based cross-enhanced protein-activated enzyme C (PKC). Action with a high antiproliferative activity, and anti-tumor, which is very suitable for the treatment of cancer. Furthermore, the high selectivity, sexual and effective inhibition of PKC has an excellent clinical effect on patients, that is, delaying or suppressing the development of the disease, and is an equally acceptable method of birth. Potential applications include a variety of solid tumors 'specifically, for example, breast cancer, colon cancer, ovarian cancer, and blood f. In addition, various other signs of PKC activity (4) can be effectively treated with these compositions, including anti-multidrug (dirty ), one of the most important problems currently used in general cancer chemotherapy and inflammatory diseases. In particular, the present invention relates to crystalline U and amorphous N-ganglars of the pharmaceutical agents disclosed herein for the treatment of such conditions or for the preparation of therapeutic agents thereof. 165155.doc •16·201247681 base-cross-aglin . The invention also relates to a method for treating a warm-blooded animal suffering from such diseases, in particular a disease of the tumor, wherein a certain amount (especially an amount having anti-colonization and anti-tumor efficacy) is crystalline π or amorphous N _Binyl-based spirulina (which is effective against related diseases) is administered to warm-blooded animals in need of such treatment. Furthermore, the present invention relates to the use of crystalline form 13 [or amorphous N-benzhydryl-spironosin for inhibiting the above PKC, or for the preparation of a pharmaceutical composition useful for treating human or animal body, in particular It is used to treat various solid tumors, such as breast cancer, colon cancer, nest cancer and blood cancer. Depending on race, age, individual symptoms, mode of administration, and clinical signs in question, an effective dose (eg, a dose of 曰 ( (9) mg, preferably 1-1000 mg, preferably 5 to 500 mg) is administered. A warm-blooded animal weighing approximately 7 kg. The present invention is also pharmaceutically acceptable in relation to a crystalline form or an amorphous formula (1) N-nodal group _ rossosporin which contains an effective amount (especially an effective amount for preventing or treating one of the diseases). A carrier for a carrier, which is suitable for topical administration; for example, for oral or rectal or parenteral administration and which may be inorganic or organic and solid or liquid. Including active ingredients (4) (iv) (eg lactose, (d) sugar, li, mannitol, sorbitol, cellulose and / or glycerin; and / or lubricants, such as vermiculite / moonstone 'hard lauric acid or its salt 'typically magnesium stearate or stearic acid Calcium acid; and / or PEG) key or gelatin capsules are especially used for oral administration. Key agents can be the same - mixture [such as Shi Ximing acid town, starch, usually corn 'wheat or starch, gelatin, Methylcellulose, buffered methylcellulose 165155.doc 201247681 Sodium and/or polyvinylpyrrolidone; and if desired, also includes disintegrants such as starch, agar, ed acid or its salts , usually sodium alginate, and/or foaming a mixture of adsorbents, colorants, flavorings and sweeteners. The pharmaceutically active compound of the present invention can be further used in the form of a preparation for parenteral administration or a soaking solution. These solutions are preferably iso-osmolar aqueous solutions or suspensions, which may be prepared prior to use, for example, as a lyophilized formulation comprising the active ingredient, either alone or together with a carrier such as mannitol. Words. The pharmaceutical material may be sterilized and/or may contain excipients such as preservatives, stabilizers, wetting agents and/or emulsifiers; solubilizers; salts which modulate osmotic pressure; and/or buffers. The pharmaceutical preparations of the invention (which may, if desired, comprise other pharmaceutically active substances, such as antimony antibiotics), are prepared in a manner known per se, for example by conventional mixing, granulating, coating, dissolving or A lyophilization process and comprises about 1% (especially from about 1% to about 20%) of a plurality of or a single active substance. The invention is further defined by reference to the following detailed description of the compounds and groups of the invention: the preparation of the invention and the use thereof. It will be apparent to those skilled in the art that many changes in the materials and methods can be practiced without departing from the spirit and scope of the invention. Subsequent examples are not intended to limit the scope of the invention to the scope defined above or the scope defined by the scope of the claims below. EXAMPLES Example 1 Purification method using spironolactone of methanesulfonic acid and triethylamine to transform into sporosmine B2 Feed cross-agrowsin B1 and ethanol into a reactor. Heat the suspension to 165155.doc 201247681 about 70 C. The broth is added to the suspension and the resulting solution is aged. Activated charcoal and Hyflo' were added and the mixture was aged. This suspension was filtered and the filter unit and filter cake were washed several times with ethanol. The clear filtered solution was transferred to another reactor and cooled to about 6 Torr. Hey. Triethylamine, which was slowly diluted in ethanol, was added to the reaction mixture. Thereafter, the suspension was cooled to about 20 ° C and aged. Sporosphingin B2 is isolated by filtration. The product was rinsed several times with ethanol. Example 2 Purification of crude staurosporin B1 to stascmarinin B2 by crystallization to feed staurosone B 1 and benzyl alcohol to a reactor. The suspension was heated to about 85 °C. Activated charcoal and Hyflo were added to the solution and the mixture was aged again. Approximately 9 (filtering this suspension at a temperature of TC. Washing the filter device and filter cake multiple times with alcohol) transferred the clear filtered solution to another reactor and cooled to about 70 ° C. The solution was Socrosin B2 suspended in ethanol is sown. Thereafter, ethanol is added to the suspension and the suspension is aged 'then' and then cooled to about 〇 ° C. The suspension is aged again for at least 2 hours at about 〇 ° C. Sporogenin B2 was isolated by filtration. The product was rinsed several times with ethanol. Example 3 Benzylated sporosporin B2 to prepare a substantially amorphous thoracic-strepostine-fed cross containing benzylic acid liver The bodily B1, ethanol and water are brought to a reactor. The reaction mixture is heated to about 70 C. The solution is filtered, and the filter device and filter cake are washed several times with ethanol. The filtered solution is transferred to another. In the reactor, and cooled to about 601. Water was slowly added to the solution. It was sown with amorphous N- > broth-cross-aglycin. The suspension was aged, then cooled to 165155.doc 201247681 About 20 ° C, and then aging. Ν · benzyl sulfhydryl - cross The gelatin was separated by filtration. The wet cake of Ν-benzyl hydrazino-spirinsin was rinsed several times with pure ethanol. The wet cake was then resuspended in a ferric device in ethanol at about 25 ° C. Under aging, then cooled to about 〇 -5 ° C, and then aged. Repeat the process several times. N-benzyl ketone - staurosporine is separated by filtration. The external temperature is about 40 ° C The wet filter cake was dried in a vacuum oven having a vacuum of about 1 〇 20 mbar for about 24 hours to obtain a product: 35-40 grams of dry cake: 27 grams of theoretical weight reduction. Yield: relative B1 100 % is a theoretical mitigation value of about 82%. The ethanol in the above reaction can be substituted with THF. Example 4 benzylated staurosporin B2 to prepare a crystalline form of ~gangyl-spironosin B4 feed containing benzylic anhydride Sporosphingin B2, pure ethanol and water to a reactor. Heat the reaction mixture to about 70. (3 and age. At the end of the reaction, use N-benzyl fluorenyl-cross suspended in pure ethanol Sporogenin crystal form 1; [sow the reaction mixture. After aging, slowly add pure ethanol and water to the suspension In the floating solution, the suspension is re-aged and then cooled to about 〇t: before the separation of N-benzyl hydrazino _ sporosporin by filtration. Rinse N-benzyl ketone-sweet sulphate with pure ethanol The filter cake was applied several times. Then it was dried in a vacuum oven with an external temperature of 60 ° C and a vacuum of about 5-30 mbar for about 16 hours to obtain a product · 39.7 G g, dry type > cake: 27 · 2 g theory Reducing the weight I. Yield. Theoretical mitigation value relative to 〇〇〇1 1〇〇〇/0 about 9ΐ.5〇/〇. Example S Preparation of crystalline form 11 of benzyl sulfhydryl copporin using benzyl alcohol and ethanol 165155.doc • 20- 201247681 Feed the crude ten-predulin and benzyl alcohol to a reactor. The mixed β was heated to 85 C, and then the solution was aged. Cool the solution to about 7 Torr. 〇, and then suspended in pure ethanol. N-V thiol _ nasosporin of the genus of the genus is sown in pure crystalline form II. Afterwards, 'add ethanol, and age the suspension, then cool to about 〇 ° C before filtration in N-blocking. The wet cake is rinsed several times with ethanol and dried. Example 6 Preparation of crystalline form 11 of bismuthinosporine with crude acetaminosine and acetic acid into a reactor using acetamidine and ethanol. The mixture was heated to about 70 ° C and then aged. This solution is sown with a pure crystalline form of N_benzylidene-spirinsin B6 suspended in pure ethanol. Thereafter, ethanol is added to the suspension, and the suspension is aged, then in Ν-benzyl The sulfhydryl-crynamicin crystal form II was cooled to about 20 ° C before separation by filtration. The wet filter cake was rinsed several times with ethanol and dried. Example 7 Preparation of N-benzyl hydrazino-craspellin using PEG Crystallized η crystals were fed into the crude Ν-benzyl aryl-sporosporin and peg to a reactor. The mixture was heated to about 90 ° C, and then the solution was aged. The solution was cooled to about 70. °C, then use N-benzyl hydrazino-spiritin B6 pure knot suspended in pure ethanol Form II seeding. After that, gradually add ethanol and water in a ratio of 1:1, and age the suspension before cooling to 20 ° C. Separation of the crystalline form of N-benzyl sulfhydryl-aspirin by filtration The wet filter cake was rinsed several times with ethanol and dried. Example 8 Preparation of N-benzyl sulfhydryl-craspellin crystal form using DMSO 165 155155.doc -21 - 201247681 Feeding N-benzyl sulfhydryl-crium The cinnamic acid and DMSO are added to a reactor. The mixture is heated to about 7 ° C and the solution is then aged, and then seeded with N-branched-Streptosporin B6 pure crystalline form II suspended in ethanol. After that, ethanol was added to the suspension, and the suspension was aged at 70 ° C, and then cooled to about 2 ° C. Separation of the crystalline form II of N-benzyl hydrazino-craspellin by filtration The suspension was aged. The product was rinsed several times with ethanol and dried. Example 9 Preparation of N-benzyl hydrazino-craspellin crystal form π feed N-benzyl hydrazino-spiritin and DMF to DMF In the reactor, heat the mixture to about 70 ° C, age the solution, and then use N-benzyl hydrazino-aspartate in ethanol The B6 pure crystalline Form II is sown. Thereafter, ethanol is added to the suspension, and the suspension is aged at about 70 ° C, and then cooled to about 20 ° C. Separation of N-benzyl fluorenyl-C. The crystalline form of the vulgaris was previously aged again. The product was rinsed several times with ethanol and dried. Example 10 Preparation of N-Benzylsulfonyl-craspellin crystal form using DMF Feeding amorphous N-benzyl Sulfhydryl-sporosporin and THF are added to a reactor. The mixture is heated to about 65 ° C, the solution is aged, and then seeded with a pure crystalline form of N-benzyl sulfhydryl-craspellin B6 in THF. The suspension was aged 'then' and then cooled to about 2 (TC) and the suspension was re-aged prior to separation of the crystalline form II of N-benzylindenyl-scaspudrin. The product was rinsed several times with THF and dried. Example 11 was prepared by using ethanol to prepare a crystalline form of N-benzylindenyl-scaspudrin. The amorphous N-flavoring-cross-aggregating agent and ethanol were fed to a reactor. The mixture was heated to about 7 (TC, aged, and then seeded with crystal form II of benzyl 165155.doc • 22· 201247681 thiol-craspellin B6 in ethanol. The suspension was aged and cooled to about 20 ° C. And separating the crystal form of N-benzyl hydrazino-craspellin by filtration. The suspension was re-aged before. The product was rinsed several times with ethanol and dried. Example 12 Spray drying experiment to obtain amorphous PKC412 10 g of PKC412 (crystalline Form II) was dissolved in 160 g of dichloromethane to prepare a feed solution. The feed was prepared and atomized into droplets, and the organic solvent was evaporated through heated nitrogen in a dry chamber and heated. The nitrogen will result in the formation of amorphous PKC 412. The inlet temperature was set to 60 ° C and the outlet temperature was set to 40 ° C. The amount of solvent remaining after spray drying was about 1% w/w. Example 13 Spray Drying Experiment Obtaining amorphous PKC412 7 g of PKC412 (crystalline Form II) was dissolved in about 400 g of THF to prepare a feed solution. The feed was prepared and atomized into droplets, and the organic solvent was passed through heated nitrogen to the dry chamber. Evaporating out, heated nitrogen will cause generation Crystalline PKC412. The inlet temperature is set to 110-120 ° C, and the outlet temperature is set to 70-80 ° C. The amount of solvent remaining after spray drying is about 3 ° / wt / weight. PKC412 is further dried under vacuum, A solvent free material was obtained. Example 14 Spray Drying Experiment to Obtain Amorphous PKC412 7 grams of PKC412 (Crystal Form II) was dissolved in about 350 grams of 70:30 halo/weight ethanol/acetic acid to prepare the feed. The solution is prepared and atomized into droplets. The organic solvent is evaporated in the dry chamber through heated nitrogen. The heated nitrogen will cause the formation of amorphous PKC412. The inlet temperature is set to 215 ° C ' and the outlet temperature is set. It is 120 ° C. The amount of residual solvent after spray drying is less than 0.5%. No further conventional drying treatment is required. 165155.doc -23- 201247681 Example 15 Crystalline form containing N-benzyl hydrazino-craspellin Tablets of II containing 100 mg of the active substance of the title are usually prepared according to the following composition: Content of the composition (mg) Active ingredient 100 Crystalline lactose 240 Avicel 80 PVPPXL 20 A highly dispersible silicone 2 Stearic acid 5 Total 447 Preparation method: The active substance and carrier material are mixed and placed on a press (Korsch EKO, punching diameter 10 mm) to compress Avicel is microcrystalline cellulose (FMC, Philadelphia, USA) PVPPXL is polyethylene based Pyrrolidone, crosslinked (Badtis, Germany)
Aerisol是二氧化石夕（Degussa，德國） 實例16含N-苄醢基-十字孢靈素的結晶形II的膠囊 含100毫克標題中所命名之化合物作為活性物質的膠囊 通常是依下列組成製備： 組合物含量 （毫克) 活性成份 100 Avicel 200 PVPPXL 15 一種高度分散的矽膠 2 硬脂酸鎂 1.5 總計 318.5 膠囊藉混合該等組份並將混合物填充至硬式明膠膠囊 (尺寸1)中而製得。 【圖式簡單說明】 165155.doc •24- 201247681 圖1顯示N-节醯基-十字孢靈素結晶形Η的X-射線# v物末繞 射圖譜。 圖2顯示N-苄醯基-十字孢靈素結晶形Π之特有的红外線 光譜。 圖3顯示N-苄醯基-十字孢靈素純非晶形的X-射線粉末繞 射圖譜。 圖4顯示N_节醯基_十字孢靈素基本上為非晶形的χ_射線 粉末繞射圖譜》 165155.doc 25·Aerisol is a dioxide dioxide (Degussa, Germany). Example 16 Capsules containing crystalline form II of N-benzyl hydrazino-craspellin containing 100 mg of the compound named in the heading as an active substance are usually prepared according to the following composition : composition content (mg) active ingredient 100 Avicel 200 PVPPXL 15 a highly dispersible silicone 2 magnesium stearate 1.5 total 318.5 capsules by mixing the components and filling the mixture into a hard gelatin capsule (size 1) . [Simple description of the diagram] 165155.doc •24- 201247681 Figure 1 shows the X-ray #v end-of-end diffraction pattern of the N-nodal thiol-cryntaxet crystalline form. Fig. 2 shows an infrared spectrum unique to the crystalline form of N-benzyl hydrazino-craspellin. Figure 3 shows a pure amorphous X-ray powder diffraction pattern of N-benzylindenyl-scaspudrin. Figure 4 shows a diffraction pattern of a substantially amorphous χ-ray powder of N_ 醯 醯 _ 十字 灵 灵 165 165155.doc 25·